Development of a shuttle vector for nocardioforms

Abstract

Construction of Escherichia coli – nocardioforms shuttle vector pAR1 was achieved using the recombinant DNA technology. Developmental strategy included following cloning and fusion steps. Firstly, insertion of kanamycin resistance cassette into the suicide vector pEcoR251 was performed. This was followed by ligations of a nocardial replicon from the pCY104 and EcoRI endonuclease from the pEcoR251, lacking ampicillin resistant determinant. New recombinant plasmid was engineered with a positive (suicide) function and a single selectable marker expressed in both Gram-positive and Gram-negative bacteria. Plasmid pAR1 was relatively small ~5.0 kb in size.